EP1185669B1 - Promotor, welcher die expression von transgenen in der ganzen pflanzen ausser dem samen erlaubt - Google Patents

Promotor, welcher die expression von transgenen in der ganzen pflanzen ausser dem samen erlaubt Download PDF

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EP1185669B1
EP1185669B1 EP00940461A EP00940461A EP1185669B1 EP 1185669 B1 EP1185669 B1 EP 1185669B1 EP 00940461 A EP00940461 A EP 00940461A EP 00940461 A EP00940461 A EP 00940461A EP 1185669 B1 EP1185669 B1 EP 1185669B1
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sequence
plant
gene
promoter
protein
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EP1185669A1 (de
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Bertrand Dubreucq
Loic Lepiniec
Michel Caboche
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Institut National de la Recherche Agronomique INRA
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells

Definitions

  • the present invention relates to the isolation and characterization of a promoter which allows the expression of transgenes in the adult plant, in order to improve the development of the plant, without the product of this transgene being present in the mature and dry seed.
  • the invention also relates to transgenic plants. comprising a gene of interest fused to said promoter sequence.
  • the specificity of expression of the transgenes introduced is essentially based on the use of promoter sequences of plants or microorganisms.
  • the search for specific promoters is therefore of importance capital for plant biotechnologies.
  • the seeds are a component important for agriculture, as seeds, but also in food or the processing industry. As such, the presence of proteins and products new to the seed can cause problems. It therefore appears interesting to be able to have an active promoter in all vegetative tissues, but ineffective in the seeds.
  • the objective underlying the present invention is to identify a promoter which would allow strong expression of a transgene in all plant tissues except in the seed.
  • promoter trapping a powerful tool for dissecting developmental processes (Topping and Lindsey 1995 for review), a been implemented.
  • This strategy is based on the use of a vector of transformation of plants having, at one of its ends, a reporter gene (GUS or GFP most often) without promoter. If integration takes place in a region coding and if the reporter gene sequence is in phase there will be fusion translational between the endogenous protein and the marker gene protein.
  • the gene trapping strategies have a major advantage over the classic insertional mutagenesis because the phenotype (expression of the reporter gene GUS) is dominant. This dominance of the phenotype (GUS) makes it possible to follow the alleles mutated to the heterozygous state. This is very interesting for the study of mutations which are lethal in the homozygous state. This approach also makes it possible to characterize a gene by its expression.
  • the present invention relates to a promoter sequence allowing the expression of a gene of interest in the tissues of a plant except in the maturing seed and in the dry seed.
  • said sequence comprising a sequence having at least 80% of identity with the sequence or a portion of the sequence of the promoter of the gene for FAH of Arabidopsis.
  • this sequence comprises a sequence having at least 80% identity with the sequence or a portion of the sequence SEQ ID No. 1.
  • % identity is meant the percentage of identical nucleotides which can easily be calculated by a person skilled in the art using a computer program for sequence comparison such as the DNASIS program (Version 2.5 for Windows; Hitachi Software Engineering Co. , Ltd, South San Francisco, CA) using the standard parameters described in the manufacturer's manual, incorporated into the description by reference.
  • the sequences and the percentages of identity can also be obtained by using internet computing resources.
  • sequence according to the invention has the following sequence or a portion of the sequence SEQ ID No. 1:
  • the invention also relates to the use of a portion of the sequence SEQ ID No. 1 for the identification of fragments capable of promoting the expression of a gene of interest in a plant except in the seed. It is thus possible to define the region minimum of the FAH gene promoter sequence to ensure expression effective. In this sense, the promoter can be modified by adding sequences such as enhancers, by deletion of non-essential and / or unwanted regions.
  • the promoter can include synthetic and / or natural sequences.
  • a primer comprising a sequence having at least 80% identity with a sequence having at least 10 consecutive nucleotides of the genomic sequence of the FAH gene of Arabidopsis (introns and exons) which is accessible to a person skilled in the art under the number AC003096 or a primer which hybridizes under stringent conditions to any coding sequence for the following SEQ ID No. 4 (Arabidopsis thaliana, fatty acid hydroxylase Fahlp):
  • the promoter sequence allowing the expression of a gene of interest in the plant tissue, except in the maturing seed and in the dry seed may also be characterized in that it comprises a sequence having at least 80% of identity with the sequence or a portion of the sequence of the promoter of the gene for FAH capable of being obtained from the process described above.
  • Another aspect of the invention relates to an expression cassette which comprises a sequence of interest fused to a sequence comprising a promoter sequence as defined above.
  • Said sequence of interest can code for an RNA, a protein or a polypeptide which protects the plant against biotic or abiotic stress.
  • the cassette can allow the co-suppression of the expression of a gene characterized in that said sequence of interest codes for a protein or a polypeptide capable of replacing the function of an endogenous protein or polypeptide.
  • the sequence of interest can also code for an antisense sequence directed against a target gene. This makes it possible, by coupling with the ectopic overexpression of a gene of interest in the seeds, to prevent any expression of this gene in other tissues. the antisense is not expressed there.
  • the cassette according to the invention can also comprise a selection marker gene, a leader sequence controlling the transit, secretion or targeting of the expression product in different organelles, a signal sequence for the termination of transcription and translation.
  • the gene of interest according to the invention can also be a gene controlling the development such as for example a gene involved in the metabolism of hormones, in signal transduction, or in cell cycle control.
  • Another aspect of the invention relates to a vector, in particular a plasmid vector, comprising an expression cassette as defined above.
  • the invention also relates to a plant cell transformed with the cassette or with a vector comprising said cassette and a plant transformation kit comprising said cassette or said vector.
  • the transfer of DNA inside plant cells may be performed by standard techniques (Plant Cell Report, 10, 595, 1992), in particular by transfer via Agrobacterium (Plant J., 1994. 6, 271), by electroporation (Nature, 1987, 327, 70), laserporation (Barlev Genetics. 1991 VI, 231), by polyethylene glycol, or by the biolistic method called "gun particle” (Nature, 1987, 327, 70).
  • Plant Cell Report, 10, 595, 1992 in particular by transfer via Agrobacterium (Plant J., 1994. 6, 271), by electroporation (Nature, 1987, 327, 70), laserporation (Barlev Genetics. 1991 VI, 231), by polyethylene glycol, or by the biolistic method called "gun particle” (Nature, 1987, 327, 70).
  • agrobacterium infiltration in planta Bechtold et al.
  • the transformation vectors carry selection markers, T-DNA borders, cloning sites, functions of replication, as well as other elements if necessary for the proper transfer of transgenes (Bouchez et al. 1993).
  • the above-mentioned publications are incorporated into the description by references.
  • the subject of the present invention is a transgenic plant capable of to be obtained by implementing the method mentioned above.
  • plant capable of being obtained any plant expressing a transgene in its tissues except in mature and dry seeds, said plant having a promoter according to the invention. Plants obtained by any process equivalent leading to the same result are also subject of the invention.
  • the list plants in which this promoter sequence can be used includes more especially plants which are useful for any industry. We can quote by example rapeseed, cruciferous, corn, soy, wheat, sunflower, pea, plants ornamental, and trees.
  • the invention relates to a plant, as defined above, which expresses in its tissues, except in seeds, a gene whose product (RNA or protein) protects the plant against biotic or abiotic stress, an antisense sequence directed against a target gene, a protein or a polypeptide capable of replacing the function of a endogenous protein or polypeptide or a coding sequence for a protein involved in the biosynthesis of metabolites or a gene controlling development such as for example a gene involved in the metabolism of hormones, in the signal transduction, or in cell cycle control.
  • RNA or protein whose product
  • an antisense sequence directed against a target gene a protein or a polypeptide capable of replacing the function of a endogenous protein or polypeptide or a coding sequence for a protein involved in the biosynthesis of metabolites or a gene controlling development such as for example a gene involved in the metabolism of hormones, in the signal transduction, or in cell cycle control.
  • the plant according to the invention can also express a protein of interest under the control of a promoter other than the promoter of the FAH gene and an antisense sequence capable to inhibit the expression of said protein of interest under the control of the promoter of the FAH gene, so that the gene of interest is only expressed in seeds.
  • the method used for the extraction of genomic DNA from Arabidopsis is inspired by that described by Doyle and Doyle (1990). The principle is based on the properties cetyltrimethylammonium bromide detergents (CTAB; Sigma Chemical Co., USA) allowing specific denaturation of protein macromolecules and polysaccharide. About 2g of plant material (seedlings grown in vitro, old 1 to 2 weeks) are finely ground in liquid nitrogen and transferred to a tube 50ml of FALCON type (Costar, USA), containing 7.5ml of extraction buffer preheated to 65 ° C. The extraction is carried out at 65 ° C for 30 minutes, with stirring regular.
  • CAB cetyltrimethylammonium bromide detergents
  • Proteins denatured by ⁇ -mercaptoethanol and CTAB buffer are then extracted into a volume of chloroform and then eliminated after centrifugation (4430g. 10min).
  • the supernatant nucleic acids are precipitated by one volume of isopropanol in the presence of 3M sodium acetate (1/10, v / v), centrifuged (7900g, 10min) then rinsed with 70% ethanol.
  • the pellet is taken up in an Eppendorf tube in 100 ⁇ l of water and the ribonucleic acids are eliminated by adding 3 ⁇ l of Rnase A at 10mg / ml (Sigma Chemical Co .. USA). DNA is deproteinized and then again precipitated with absolute ethanol. After centrifugation in an Eppendorf tube, the pellet is washed. dried, taken up in 50 to 100 ⁇ l of water and stored at -20 ° C. before analyzes.
  • the promoter sequence was amplified using PCR technology which is a known technique (Sambrook et al. 1989). Primers corresponding to the 5 '(upper) and 3' (lower) part of the promoter sequence were derived from the genomic sequence of BAC T29F13 (AC003096) (See FIG. 1). Genomic DNA from a wild-type line (Ler) was used as a template for amplification of the promoter part.
  • the amplification reactions were carried out on a thermocycler (MJ Research PTC100 -96), in 0.2 ml tubes (Prolabo) containing the following mixture: 1 ⁇ l (10ng) DNA, 2 ⁇ l Buffer 10 x (BRL), 2 ⁇ l MgCl2 25mM, 0.8 ⁇ l dNTP 5mM, 1 ⁇ l Primer 1 (10 pmole/ ⁇ l), 1 ⁇ l Primer 2 (10 pmole/ ⁇ l), 0.5 ⁇ l (1U) Taq DNA polymerase (5U / ⁇ l), H2O qs 20 ⁇ l
  • the genotypes of bacteria used for carrying out the experiments are:
  • E. Coli strain DH12S ( ⁇ 80 dlacZ ⁇ M15 mcrA ⁇ (mrr-hsdRMS-mcrBC) araD139 ⁇ (ara, leu) 7697 ⁇ lacX74 galU galK rpsL deoR nupG recAl / F'proAB + laclq Z ⁇ M15).
  • the transformation of bacteria E. coli strain DH12S by a recombinant plasmid is performed by electroporation (Potter, 1993).
  • electroporation tank (1ml, width 0.1cm)
  • 2 ⁇ l of the ligation reaction are mixed with 50 ⁇ l of bacteria thawed and kept in ice.
  • the tank is then placed in a electroporator (Gene Pulser II System: BIO-RAD, FRANCE) and a voltage of 1.25kV is applied for a time which is a function of the resistance (200 ⁇ ) and the capacity (25 ⁇ F) of the circuit.
  • BIO-RAD BIO-RAD
  • FRANCE electroporator
  • a voltage of 1.25kV is applied for a time which is a function of the resistance (200 ⁇ ) and the capacity (25 ⁇ F) of the circuit.
  • One ml of SOC medium is added to promote growth bacteria and the whole is incubated in a 10 ml tube for 2 hours at 37 ° C.
  • the transformed bacteria are then spread on boxes containing solid LB medium supplemented with the appropriate antibiotic and incubated at 37 ° C overnight.
  • the selection of bacteria transformed by the recombinant pMeca plasmid is made with 0.04 mg / ml of ampicillin in the presence of 0.2mg / ml of X-Gal and 0.05mg / ml of IPTG.
  • the bacteria are selected on an LB medium with the appropriate antibiotic for a final concentration of 0.04mg / ml.
  • the sequence of the gene in question was obtained using sequences from the systematic sequencing of the Arabidopsis thaliana genome and is found to be localized on the BACT29F13.
  • An expressed sequence (EST TAI234) has been identified in databases and appears to correspond to a full length sequence of mRNA FAH. This allowed the identification of the untranslated 5 'transcribed sequence and of the intended location of the promoter sequence.
  • the intron / exon structure has been deduced, at the level of the untranslated transcribed part, the alignment of the BAC with the EST TAI234 ( Figure 1).
  • the promoter was amplified by PCR from the primer pFAH / upper and the primer A1, placed in the untranslated transcribed part 5 ′ (FIG. 2).
  • a study of the sequence has shown that the amplified sequence has a putative TATA box at -100bp from the site of suspected transcription initiation (based on full-length cDNA) and a box CCAAT at -190bp from the same start of transcription.
  • the amplified PCR fragment (932pb) was cloned into a vector pGEM-T (PROMEGA), sequence, then introduced in a binary vector (pBI101, Clontech) containing a GUS reporter gene without promoter (Figure 3). This construction was then introduced by transformation in planta, via Agrobacterium, in wild type plants (Ws ecotype). Thirteen primary transformants were obtained, which were tested for their GUS activity during their development.
  • the expression is strong from 20 hours after the start of imbibition in the embryo. During development, expression is strong in all tissues, with a certain preference for vascular tissues.
  • GUS reporter gene expression profile cotyledons adult leaf roots flower silicic germinating seeds Dry seeds 1 ++ +++ ++ ++ +++ +++ - 2 +++ +++ ++ ++ +++ +++ - 3 +++ +++ ++ ++ nd ++ - 4 +++ +++ ++ ++ ++ nd ++ - 5 +++ +++ ++ ++ nd +++ - 6 + +++ ++ ++ nd +++ - 7 +++ +++ ++ ++ + - 8 ++ +++ ++ ++ +++ +++ +++ - 9 +++ +++ ++ ++ ++ - - -
  • the expression of the marker confirms the functionality of the promoter and its specificity.
  • This type of promoter is therefore of great interest for applications biotechnology, such as the expression of an anti-insect toxin (type Bt) in plants and the expression of any transgene making it possible to improve, so quantitative or qualitative, the development and growth of the plant without the protein encoded by the transgene is present in the seed.
  • type Bt anti-insect toxin

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Claims (23)

  1. Promotorsequenz, welche die Expression eines Gens von Interesse in den Geweben einer Pflanze außer in dem in Reifung befindlichen Samenkorn und in dem trockenen Samenkorn erlaubt, dadurch gekennzeichnet, dass sie eine Sequenz umfasst, welche wenigstens 80% Identität mit der Sequenz SEQ ID No. 1 des Promotors des Gens der FAH von Arabidopsis aufweist.
  2. Sequenz nach Anspruch 1, dadurch gekennzeichnet, dass sie eine Sequenz umfasst, die unter Bedingungen hoher Stringenz mit der Sequenz SEQ ID No. 1 hybridisiert.
  3. Sequenz nach Anspruch 1, dadurch gekennzeichnet, dass sie die Sequenz SEQ ID No. 1 umfasst.
  4. Verfahren, welches die Isolierung und Charakterisierung des Promotors des Gens der FAH in Pflanzen, ermöglicht, welches die folgenden Schritte umfasst:
    a) Verwendung eines Primers, welcher eine Sequenz umfasst, welche wenigstens 80% Identität mit einer Sequenz, welche wenigstens 10 aufeinanderfolgende Nukleotide der Sequenz SEQ ID No. 5 oder einer komplementären Sequenz aufweist, aufweist, eines Primers, der unter Bedingungen hoher Stringenz mit einer jeglichen SEQ ID No. 4 kodierenden Sequenz oder einer Sequenz, welche wenigstens 80% Identität mit einer Sequenz, die wenigstens 10 aufeinanderfolgende Nukleotide der genomischen Sequenz des Gens der FAH von Arabidopsis, welche unter der Nummer AC003096 zugänglich ist, oder einer komplementären Sequenz aufweist, aufweist, hybridisiert, für die Isolierung und/oder Amplifizierung der Sequenz stromaufwärts von dem 5'-Ende des Gens der FAH,
    b) Klonierung und Sequenzierung der in Schritt a) erhaltenen Sequenz.
  5. Verwendung einer Sequenz nach einem der Ansprüche 1 bis 3 für die Identifizierung von Fragmenten der Sequenz SEQ ID No.1, welche die Expression eines Gens von Interesse in den Geweben einer Pf0lanze außer in dem in Reifung befindlichen Samenkorn und in dem trockenen Samenkorn erlauben.
  6. Expressionskassette, dadurch gekennzeichnet, dass sie eine Sequenz von Interesse fusioniert mit einer Sequenz, welche eine Promotorsequenz nach einem der Ansprüche 1 bis 3 umfasst, umfasst.
  7. Expressionskassette nach Anspruch 6, dadurch gekennzeichnet, dass die Sequenz von Interesse eine RNA, ein Protein oder ein Polypeptid, welche bzw. welches die Pflanze gegen einen biotischen oder abiotischen Stress schützt oder welche bzw. welches an der Entwicklung, insbesondere am Metabolismus der Hormone, an der Transduktion von Signalen oder an der Kontrolle des Zellzyklus, beteiligt ist, kodiert.
  8. Expressionskassette nach Anspruch 6, welche die Cosuppression eines Gens erlaubt, dadurch gekennzeichnet, dass die Sequenz von Interesse ein Protein oder ein Polypeptid kodiert, welches in der Lage ist, die Funktion eines endogenen Proteins oder Polypeptids zu ersetzen.
  9. Expressionskassette nach Anspruch 6, dadurch gekennzeichnet, dass die Sequenz von Interesse eine Antisinn-Sequenz, welche gegen ein Zielgen gerichtet ist, kodiert.
  10. Expressionskassette nach Anspruch 6, dadurch gekennzeichnet, dass die Sequenz von Interesse ein Enzym, welches an der Produktion von Metaboliten durch eine Pflanze beteiligt ist, kodiert.
  11. Vektor, welcher eine Expressionskassette nach einem der Ansprüche 6 bis 9 umfasst.
  12. Mit einer Kassette nach einem der Ansprüche 6 bis 9 oder einem Vektor nach Anspruch 11 transformierte Pflanzenzelle.
  13. Kit zur Transformation von Pflanzen, welcher eine Kassette nach einem der Ansprüche 6 bis 9 oder einen Vektor nach Anspruch 11 umfasst.
  14. Verfahren zur Erzeugung von transgenen Pflanzen, in welchen ein Gen von Interesse in allen Geweben außer in dem in Reifung befindlichen Samenkorn und in dem trokkenen Samenkorn exprimiert wird, dadurch gekennzeichnet, dass es die folgenden Schritte umfasst:
    a) Transfer einer Kassette nach einem der Ansprüche 6 bis 9 oder eines Vektors nach Anspruch 11 in Zellen von Pflanzen,
    b) Kultivierung der in Schritt a) erhaltenen transformierten Zellen derart, dass die transgenen Pflanzen erhalten werden.
  15. Verfahren nach Anspruch 14, dadurch gekennzeichnet, dass die Zellen unter den embryonalen Zellen, welche von einem unreifen Embryo stammen, ausgewählt werden.
  16. Verfahren nach einem der Ansprüche 14 und 15, dadurch gekennzeichnet, dass der Transfer unter Verwendung von Agrobacterium, vorzugsweise Agrobacterium tumefaciens ausgeführt wird.
  17. Transgene Pflanze, dadurch gekennzeichnet, dass sie eine Kassette nach den Ansprüchen 6 bis 10 umfasst.
  18. Pflanze nach Anspruch 17, dadurch gekennzeichnet, dass sie in ihren Geweben außer in den reifen und trockenen Samenkörnern eine RNA, eine Antisinn-Sequenz, welche gegen ein Zielgen gerichtet ist, exprimiert.
  19. Pflanze nach Anspruch 17, dadurch gekennzeichnet, dass sie in ihren Geweben außer in den reifen und trockenen Samenkörnern eine RNA, ein Protein oder ein Polypeptid, welche in der Lage sind, die Funktion eines endogenen Proteins oder Polypeptids zu ersetzen, exprimiert.
  20. Pflanze nach Anspruch 17, dadurch gekennzeichnet, dass sie ein Protein von Interesse unter der Kontrolle eines anderen Promotors als des Promotors des Gens der FAH und eine Antisinn-Sequenz, welche in der Lage ist, die Expression des Proteins von Interesse zu inhibieren, unter der Kontrolle des Promotors des Gens der FAH exprimiert derart, dass das Protein von Interesse lediglich in den Samenkörnern exprimiert wird.
  21. Pflanze nach Anspruch 17, dadurch gekennzeichnet, dass sie in ihren Geweben außer in den reifen und trockenen Samenkörnern eine Sequenz, welche ein Protein, das an der Biosynthese von Metaboliten beteiligt ist, ein Protein oder ein Polypeptid, welches die Pflanze gegen einen biotischen oder abiotischen Stress schützt, ein Protein, welches die Entwicklung, insbesondere im Rahmen des Metabolismus der Hormone, im Rahmen der Transduktion von Signalen oder im Rahmen der Kontrolle des Zellzyklus, kontrolliert, kodiert, exprimiert.
  22. Pflanze nach einem der Ansprüche 17 bis 21, dadurch gekennzeichnet, dass sie insbesondere unter Raps, den Kreuzblütlern, Mais, Soja, Getreide (Weizen), Sonnenblume, Erbse, den Zierpflanzen und den Bäumen ausgewählt wird.
  23. Samenkörner, welche ausgehend von einer transgenen Pflanze nach einem der Ansprüche 17 bis 22 erhalten werden, dadurch gekennzeichnet, dass sie das Expressionsprodukt des Transgens nicht enthalten, wobei die Samenkörner eine Kassette nach einem der Ansprüche 6 bis 10 oder einen Vektor nach Anspruch 11 umfassen.
EP00940461A 1999-06-10 2000-06-08 Promotor, welcher die expression von transgenen in der ganzen pflanzen ausser dem samen erlaubt Expired - Lifetime EP1185669B1 (de)

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FR9907362A FR2794772B1 (fr) 1999-06-10 1999-06-10 Promoteur permettant l'expression de transgenes dans toute la plante hormis dans la graine
FR9907362 1999-06-10
PCT/FR2000/001574 WO2000077223A1 (fr) 1999-06-10 2000-06-08 Promoteur permettant l'expression de transgenes dans toute la plante hormis dans la graine

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CN1555413A (zh) * 2001-07-13 2004-12-15 �ȷ�߼����ֹ��ʹɷ����޹�˾ 维管组织优选的启动子
FR2828210B1 (fr) * 2001-08-01 2004-08-06 Agronomique Inst Nat Rech Acide nucleique regulateur permettant l'expression d'un polynucleotide d'interet specifiquement dans l'endothelium d'une graine de plante, et ses applications
EP2166098B1 (de) * 2004-10-05 2013-11-06 SunGene GmbH Konstitutive Expressionskassetten zur Regulation der Pflanzenexpression
BRPI0608829A2 (pt) 2005-04-19 2011-03-15 Basf Plant Science Gmbh método para a expressão transgênica com especificidade intensificada em uma planta, uso de um construto de ácido nucleico quimérico, seqüência de ribonucleotìdeo quimérica, construto de expressão, vetor de expressão, organismo não-humano ou célula transformada, semente transformada, e, preparação farmacêutica
WO2006131490A1 (en) * 2005-06-09 2006-12-14 Basf Plant Science Gmbh Expression cassettes for seed-preferential expression in plants
US20180100162A1 (en) 2016-10-11 2018-04-12 Dow Agrosciences Llc Modulation of Transgene Expression in Plants

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US6310194B1 (en) * 1994-09-26 2001-10-30 Carnegie Institution Of Washington Plant fatty acid hydroxylases
GB9706381D0 (en) * 1997-03-27 1997-05-14 Cambridge Advanced Tech Improvements relating to the specificity of gene expression

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FR2794772B1 (fr) 2001-09-21
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DE60013123T2 (de) 2005-08-11
CA2376878A1 (fr) 2000-12-21
EP1185669A1 (de) 2002-03-13
WO2000077223A1 (fr) 2000-12-21
ATE274062T1 (de) 2004-09-15
DE60013123D1 (de) 2004-09-23
ES2223533T3 (es) 2005-03-01
PT1185669E (pt) 2004-12-31

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